Methods for decellularizing animal tissue and bioinks derived therefrom

Abstract

A method of decellularizing in an animal tissue includes digesting the animal tissue; treating the animal tissue with a surfactant: treating the animal tissue with at least one zwitterionic detergent to form a decellularized animal tissue; and treating the decellularized animal tissue with at least one advanced glycation end-product (AGE) inhibitor to reduce AGE crosslinking in the decellularized animal tissue. The decellularized animal tissue may be ground into a powder and dissolved in a digest solution to form a bioink composition useful for 3D printing an in vitro model of healthy or diseased tissue.

We claim: 
     
         1 . A method of decellularizing an animal tissue comprising:
 digesting the animal tissue;   treating the animal tissue with a surfactant;   treating the animal tissue with at least one zwitterionic detergent to form a decellularized animal tissue; and   treating the decellularized animal tissue with at least one advanced glycation end-product (AGE) inhibitor to reduce AGE crosslinking in the decellularized animal tissue.   
     
     
         2 . The method of  claim 1 , wherein the animal tissue is skeletal muscle tissue. 
     
     
         3 . The method of  claim 1 , wherein the animal tissue is a diseased tissue having enhanced AGE crosslinking in an extracellular matrix as compared to healthy animal tissue. 
     
     
         4 . The method of  claim 3 , wherein the enhanced AGE crosslinking is caused by aging, diabetes, muscular dystrophy, disuse atrophy, denervation atrophy, trauma, polymyositis, or amyotrophic lateral sclerosis. 
     
     
         5 . The method of  claim 1 , wherein the surfactant is selected from the group consisting of t-Octylphenoxypolyethoxyethanol, polysorbate 20, and polysorbate 80. 
     
     
         6 . The method of  claim 1 , wherein the at least one zwitterionic detergent comprises 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS). 
     
     
         7 . The method of  claim 1 , wherein the at least one zwitterionic detergent comprises sulfobetaine-16 (SB-16). 
     
     
         8 . The method of  claim 1 , wherein the at least one zwitterionic detergent comprises CHAPS and SB-16. 
     
     
         9 . The method of  claim 1 , wherein the at least one AGE inhibitor is selected from the group consisting of alargebrium chloride 711 (ALT-711), ALT-946, ALT-462, ALT-486, ALT-TRC4186, and aminoguanidine. 
     
     
         10 . The method of  claim 1 , wherein the at least one AGE inhibitor is ALT-711. 
     
     
         11 . The method of  claim 10 , wherein the decellularized animal tissue is treated with 0.25-30% w/v of ALT-711. 
     
     
         12 . The method of  claim 1 , wherein the animal tissue is human tissue. 
     
     
         13 . A method of manufacturing a bioink composition comprising:
 digesting an animal tissue;   treating the animal tissue with a surfactant;   treating the animal tissue with at least one zwitterionic detergent to form a decellularized animal tissue;   treating the decellularized animal tissue with at least one advanced glycation end-product (AGE) inhibitor to reduce AGE crosslinking in the decellularized animal tissue;   grinding the decellularized animal tissue to form decellularized tissue matrix material powder, and   dissolving the powder in a digest solution to form the bioink composition.   
     
     
         14 . The method of  claim 13 , wherein the grinding step is performed at—(150-200)° C. 
     
     
         15 . The method of  claim 13 , wherein the digest solution comprises pepsin. 
     
     
         16 . A method of manufacturing an in vitro model of an animal tissue comprising:
 manufacturing a bioink composition according to  claim 13 ; and   printing the bioink composition to form the in vitro model of the animal tissue.   
     
     
         17 . The method of  claim 16 , wherein the animal tissue is a healthy tissue. 
     
     
         18 . The method of  claim 16 , wherein the animal tissue is a diseased tissue having enhanced AGE crosslinking in an extracellular matrix as compared to healthy animal tissue. 
     
     
         19 . The method of  claim 18 , wherein the enhanced AGE crosslinking is caused by aging, diabetes, muscular dystrophy, disuse atrophy, denervation atrophy, trauma, polymyositis, or amyotrophic lateral sclerosis 
     
     
         20 . A bioink for physiological 3D-printing, wherein the bioink is made by the method of  claim 13 .